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建立了茶树叶片中可溶性糖的高效液相色谱一蒸发光散射检测(HPLC-ELSD)的测定方法。以AgilentZorbax carbohydrate柱为分离柱,流动相为乙腈-水(75:25,V/V),流速1.0 mL.min-1,柱温30℃,上样量10μL。蒸发光散射检测器漂移管温度85℃,雾化器温度36℃。通过该方法测定越冬期间皖茶91、乌牛早、平阳特早和龙井43叶片中果糖、葡萄糖和蔗糖的含量变化。结果表明,越冬期间3种可溶性糖含量的变化幅度一致:皖茶91>龙井43>平阳特早>乌牛早。在茶树叶片中的含量均随着气温的降低呈现增长趋势,且具有较好的规律性。蔗糖作为茶树叶片中可溶性糖的主要成分,用HPLC-ELSD法可以方便、快速、准确的检测出来。因此低温胁迫下通过HPLC-ELSD法测定茶树叶片中蔗糖含量的变化,可作为鉴定茶树品种抗寒性的一种新方法。
A method for the determination of soluble sugar in tea leaves by high performance liquid chromatography-evaporative light scattering (HPLC-ELSD) was established. The Agilent Zorbax carbohydrate column was used as the separation column. The mobile phase consisted of acetonitrile-water (75:25, V / V), flow rate 1.0 mL · min-1 and the column temperature was 30 ℃. Evaporative light scattering detector drift tube temperature 85 ℃, nebulizer temperature 36 ℃. The contents of fructose, glucose and sucrose in tea leaves 91, Wuniuzao, Pingyangtejian and Longjing 43 during winter were determined by this method. The results showed that the changes of three soluble sugar contents during the winter were consistent: Anhui Tea 91> Longjing 43> Pingyangtezao> Wuniuzao. The content of tea leaves showed an increasing trend with the decrease of temperature, and had a good regularity. As the main component of soluble sugar in tea leaves, sucrose can be detected conveniently, rapidly and accurately by HPLC-ELSD. Therefore, the changes of sucrose content in tea leaves by HPLC-ELSD under low temperature stress can be used as a new method to identify the cold resistance of tea varieties.